CN105287692A - Dalbergia benthami prain extractive, and preparation method and application thereof - Google Patents

Abstract

The invention discloses an extract of Dalbergia japonica, which is obtained by extracting the stem part of Dalbergia japonica by using ethyl acetate to reflux. The invention discloses a preparation method of the extract of Dalbergia liangyue. The stem of Dalbergia liangyue is crushed to obtain powder, and the powder is reflux extracted with ethyl acetate at 80°C for 2-4 times, each time 2-4 times. 3h, combine the filtrates extracted by reflux to obtain an extract, remove the ethyl acetate in the extract, and dry to obtain an extract of Dalbergia japonica. The invention discloses the application of the extract of Dalbergia japonica in the preparation of anti-inflammatory and antibacterial pharmaceutical preparations. The Dalbergia japonica extract of the invention has obvious anti-inflammatory and antibacterial effects, and can replace the application of other allergenic drugs in anti-inflammatory and antibacterial, thereby providing a new approach.

Description

Dalbergia japonica extract and its preparation method and application

technical field

The invention relates to an extract of Dalbergia japonica from Guangdong and its preparation method and application.

Background technique

Inflammation (inflammation) refers to a living tissue with a vascular system, which is a pathological process that mainly defends against damage caused by various inflammatory factors. The basic pathological changes are deterioration, exudation and hyperplasia.

Many Chinese herbal medicines and natural medicines have the effect of inhibiting and killing bacteria, and their anti-inflammatory effects have been paid more and more attention. Zhuang medicine Dalbergiabenthami Prain, also known as: banana vine hemp, Liangyue sandalwood, vines, born in sparse forests and bushes, distributed in Guangxi, Guangdong, Hainan and other places, the medicinal part is the stem, also known as vine spring Caulis Dalbergiabenthami, which can be used to promote menstruation and activate blood circulation, mainly treats symptoms such as bruises, muscle and bone pain, and irregular menstruation caused by qi stagnation and blood stasis. The lack of scientific basis has greatly affected the development and clinical application of Dalbergia japonica, and there is no description of the application of Dalbergia japonica extract in anti-inflammatory and antibacterial.

Contents of the invention

It is an object of the present invention to solve at least the above-mentioned problems and to provide at least the advantages which will be described later.

Another purpose of the present invention is to provide an extract of Dalbergia japonica, which is obtained by extracting its stem with ethyl acetate reflux and then drying. The extract of Dalbergia japonica of the present invention has obvious anti-inflammatory and antibacterial effects.

In order to achieve these objects and other advantages according to the present invention, an extract of Dalbergia liangyue is provided, which is obtained by extracting the stem of Dalbergia liangyue by reflux with ethyl acetate.

A preparation method of the extract of Dalbergia japonica, the stems of Dalbergia japonica are crushed to obtain powder, and the powder is reflux extracted with ethyl acetate at 80°C for 2-4 times, each time for 2-3 hours, Combine the filtrates extracted under reflux to obtain an extract, remove the ethyl acetate in the extract, and dry to obtain an extract of Dalbergia japonica.

Preferably, in the preparation method of the Dalbergia japonica extract, the powder is refluxed and extracted with ethyl acetate for 3 times, each time for 2.5 hours.

Preferably, in the preparation method of the Dalbergia japonica extract, the drying is vacuum drying, and the drying temperature is lower than 5°C.

Preferably, in the preparation method of the Dalbergia japonica extract, the amount of ethyl acetate is 5-10 times of the weight of the extract to be extracted in each reflux extraction.

An application of the extract of Dalbergia japonica, the application of the extract of Dalbergia japonica in the preparation of anti-inflammatory and antibacterial pharmaceutical preparations.

Preferably, for the application of the Dalbergia luteum extract, the pharmaceutical preparation comprises an effective amount of the Dalbergia luteum extract and pharmaceutically acceptable auxiliary materials.

Preferably, for the application of the Dalbergia japonica extract, the pharmaceutical preparation is an oil, an ointment, a cream, a paste or an ointment.

The present invention at least includes the following beneficial effects: the preparation method of Dalbergia japonica is simple, easy to obtain, has significant anti-inflammatory and antibacterial effects, and has a broad market scenario for enhancing anti-inflammatory and antibacterial effects in other products, and can replace The application of penicillin and berberine in medicine can be applied to users who are allergic to penicillin or berberine, thereby providing a new way to make up for the inability of consumers to use some anti-inflammatory and antibacterial drugs due to allergies.

Other advantages, objectives and features of the present invention will partly be embodied through the following descriptions, and partly will be understood by those skilled in the art through the study and practice of the present invention.

detailed description

The present invention will be further described in detail below in conjunction with specific embodiments, so that those skilled in the art can implement it with reference to the description.

It should be noted that the experimental methods described in the following embodiments, unless otherwise specified, are conventional methods, and the reagents and materials, unless otherwise specified, can be obtained from commercial sources.

Raw material source: Guangdong Dalbergia, identified as Dalbergia benthami Prain (Dalbergiabenthami Prain).

An extract of Dalbergia liangyue is obtained by extracting the stem of Dalbergia liangyue by reflux with ethyl acetate.

Example 1:

A method for preparing the extract of Dalbergia japonica. The stems of Dalbergia japonica are taken, dried naturally, and pulverized to obtain powder, and the powder is reflux extracted twice with ethyl acetate at 80°C, each time for 2 hours , the filtrate that merges reflux extraction obtains extracting liquid, removes the ethyl acetate in described extracting liquid, promptly obtains two Guangdong Dalbergia extracts after drying, wherein, the consumption of the ethyl acetate that extracts each time is the weight of the extract to be extracted 5 times.

Example 2:

A method for preparing the extract of Dalbergia japonica. The stems of Dalbergia japonica are taken, dried naturally, and pulverized to obtain powder, and the powder is reflux extracted with ethyl acetate at 80° C. for 3 times, each time 2.5 h, combining the filtrates extracted by reflux to obtain an extract, removing ethyl acetate in the extract, and drying to obtain an extract of Dalbergia japonica. Wherein, the amount of ethyl acetate used for each extraction is 8 times of the weight of the extract to be extracted.

Example 3:

A method for preparing the extract of Dalbergia japonica. The stems of Dalbergia japonica are taken, dried naturally, and pulverized to obtain powder, and the powder is reflux extracted with ethyl acetate at 80°C for 4 times, each time for 3 hours , combined the filtrates extracted by reflux to obtain an extract, removed the ethyl acetate in the extract, and dried to obtain an extract of Dalbergia japonica. Wherein, the amount of ethyl acetate used for each extraction is 10 times of the weight of the extract to be extracted.

Example 4:

Dried the stems of Dalbergia japonica in the shade, crushed and weighed 50g, refluxed 3 times with 400mL ethyl acetate at 80°C for 3 hours each time, combined the filtrate three times, put the filtrate in a rotary evaporator and distilled under reduced pressure, recovered the solvent, and left The product is vacuum-dried at low temperature to dryness to obtain the ethyl acetate extract (EAE) of the stem part of Dalbergia japonica, that is, the extract of Dalbergia japonica of the present invention, and the yield is generally 2.0%-2.8%.

Example 5:

The Dalbergia japonica extract of Example 4 was crushed through a 200-mesh sieve, and an appropriate amount of oil phase base, consistency regulator and emulsifier was heated and melted at about 75°C to prepare an oil phase, and an appropriate amount of moisturizer, stabilizer and water were mixed Put it in a container and heat it at 70-75°C, mix well to get the water phase, mix the oil phase with the water phase, mix it with ultrasonic, add the Dalbergia chinensis extract at 50-60°C while stirring, and cool down That is to obtain preparations in different dosage forms such as oil, ointment, cream, paste or ointment. The specific dosage form is mainly based on the needs of the applicable products. Generally, the content of Dalbergia Dalbergia extract per milliliter of liquid preparation or per gram of paste preparation is about 20-200mg. For example, if you use it on the face, you should follow the principle of small amount and multiple times.

Among them, the oil phase base includes but not limited to stearic acid, paraffin, beeswax, higher alcohol, etc., the consistency regulator includes but not limited to vaseline, liquid paraffin, vegetable oil, etc., and the emulsifier includes but not limited to soap emulsifier, polyoxyethylene Ether derivatives; humectants include but not limited to glycerin, sorbitol, propylene glycol, etc.; stabilizers can be preferably used but not limited to disodium edetate.

In order to further illustrate the anti-inflammatory and antibacterial effects of the Dalbergia japonica extract of the present invention, the present invention evaluates the anti-inflammatory activity of the Dalbergia japonica extract in vitro for the first time by establishing an acute inflammation model of mouse ear shell swelling, The antibacterial activity of Dalbergia japonica extracts was studied by filter paper diffusion method and test tube method (MIC).

1. Research on anti-inflammatory effect of effective extract in vitro

(1) Xylene-induced mouse auricle swelling experiment

40 Kunming mice with standard body weight were randomly divided into four groups (blank group, large dose of Dalbergia spp., medium dose of Dalbergia spp., and small dose of Dalbergia spp.). Xylene was evenly applied to the front and rear sides of the left ear shell of the mice to induce inflammation, 0.04ml/mouse. After the successful modeling, it was applied to the inflammation site of the left ear of the mouse respectively, and the right ear of the mouse was left untreated. After 4 hours, the animals were sacrificed by dislocation, and both ears were immediately cut along the base line of the auricles, and the ear pieces of the same part were taken with a hole punch with a diameter of 8 mm, weighed on a ten-thousandth analytical balance, and the weight of the left ear piece was subtracted from the weight of the right ear piece It is the degree of swelling, which is the degree of swelling by subtracting the weight of the ear piece on the side of the inflammation caused by the weight of the ear piece on the control side, and observe the change of the degree of swelling before and after the treatment.

Ear swelling degree = ear weight after application - ear weight before application

(2) Data processing: SPSS13.0 homogeneity of variance test.

Table 1 is the effect of the Dalbergia japonica extract measured after 4h after the application on the ear swelling of mice, as can be seen, along with the increase in dosage of the Dalbergia japonica extract, the swelling suppression rate significantly increases, indicating that The anti-inflammatory effect of Liangyue Dalbergia sandalwood extract is remarkable, and the anti-inflammatory effect will be better if you insist on using it 3 times a day.

Table 1 Effect of Dalbergia japonica extracts on mouse ear swelling

2. In vitro antibacterial activity of Dalbergia japonica extracts

(1) Bacterial solution preparation: five strains of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and Bacillus typhi were respectively marked on the MH agar plate, cultured at 37°C for 24 hours; the inoculation loops were picked separately A single colony of each strain was inoculated in MH broth and cultured at 37°C for 24 hours; the concentration of the five strains was adjusted by McFarland turbidimetry, and the final concentration was 1.5×106 CFU/mol.

(2) Filter paper diffusion method: Take 0.1ml of the bacterial suspension and spread it evenly on the plate culture medium (about 15mL) with a sterile coating rod, and put a filter paper disc with a diameter of 9mm after sterilization in the liquid medicine (concentration: 250μg/ mL) for 20 minutes, take it out, stick it on the flat plate at equal distances, put 4 pieces in each plate, and make 3 plates for each strain. Sterile water was used as a blank control, penicillin was used as a positive control for Gram-positive bacteria, and berberine was used as a positive control for Gram-negative bacteria. The antibacterial effects of Dalbergia japonica extracts on five species of bacteria are shown in Table 2.

Table 2 The antibacterial effect of Dalbergia japonica extracts on five kinds of bacteria

Note: "----" means not determined.

It can be seen from Table 2 that the extract of Dalbergia japonica has obvious inhibitory effect on Bacillus subtilis, the size of the inhibition zone is 12mm, and it has obvious inhibitory effect on Staphylococcus aureus, the size of the inhibition zone is 13mm.

(3) Test tube method (MIC): As shown in Table 3, the minimum inhibitory concentration was measured by the test tube double dilution method. 9 sterile test tubes were taken, and 1 mL of drug-containing broth was added to the first tube as a negative control. The drug concentration was 400mg/mL, add 1mL of each broth containing drug solution to tubes 2-8, in which the drug concentration is sequentially diluted twice, and add broth without drug solution to tube 9 as a positive control; then in tubes 1-8 Each of the 8 tubes was added with 50ul of the test bacteria solution cultured for 8h, shaken well, cultured at 37°C for 24h, and observed the results. The test bacteria solution here is the bacteria solution of Staphylococcus aureus and Bacillus subtilis, wherein the concentration of these two bacteria is 1.5×106 CFU/mol.

Table 3 Minimum Inhibitory Concentration Determination Test

Based on the above experiments, the applicant of the present invention obtained that the minimum inhibitory concentrations of the extracts of Dalbergia japonica from Guangdong to Bacillus subtilis and Staphylococcus aureus were 100 and 50 mg/mL, respectively.

In order to further illustrate the curative effect of the Dalbergia japonica extract of the present invention, enumerate some cases:

Typical cases:

1) Mr. Zhang, 50 years old, rheumatoid arthritis, red and swollen joints, pain in walking, smeared the external cream that adopts the Dalbergia japonica extract of the present invention to prepare (wherein, the extract of Dalbergia japonica in every 1g cream for external use The content is 50mg), 3 times on the 1st, use 7 days, redness and swelling symptom weakens, continue to use again 7 days, walking pain relief, visit after 2 months, pain obviously alleviates, insists on using external cream of the present invention for 8 months Afterwards, a follow-up visit showed that there was basically no difference in walking, and it was no different from ordinary people. After 12 months of persistent use, the pain disappeared. After 18 months of continuous use, the patient returned to visit and the condition did not repeat.

2) Ms. Jian, 30 years old, used certain cosmetics to cause facial allergies, redness, inflammation, and macules, and used the oil preparation prepared from the extract of Dalbergia japonica of the present invention (wherein every 1mL of oil preparation contained two yellow sandalwood The content of sandalwood extract is 20mg), 3 times a day, and after 3 days of use, the allergic symptoms are relieved and obviously alleviated. After continuing to use for 4 days, the macule disappears and the inflammation subsides.

Although the embodiment of the present invention has been disclosed as above, it is not limited to the use listed in the specification and implementation, it can be applied to various fields suitable for the present invention, and it can be easily understood by those skilled in the art Therefore, the invention is not limited to the specific details and embodiments shown and described herein without departing from the general concept defined by the claims and their equivalents.

Claims (8)

1. a Lignum Dalbergiae benthamii extract, is characterized in that, is the extract that the stem adopting ethyl acetate backflow to extract Lignum Dalbergiae benthamii obtains.

2. the preparation method of a Lignum Dalbergiae benthamii extract as claimed in claim 1, it is characterized in that, Lignum Dalbergiae benthamii is got its stem and pulverize to obtain powder, by described powder ethyl acetate reflux, extract, 2-4 time at 80 DEG C, each 2-3h, the filtrate merging reflux, extract, obtains extracting solution, removes the ethyl acetate in described extracting solution, namely obtains Lignum Dalbergiae benthamii extract after drying.

3. the preparation method of Lignum Dalbergiae benthamii extract as claimed in claim 2, is characterized in that, described powder ethyl acetate backflow is extracted 3 times, each 2.5h.

4. the preparation method of Lignum Dalbergiae benthamii extract as claimed in claim 2, it is characterized in that, described drying is vacuum drying, and baking temperature is lower than 5 DEG C.

5. the preparation method of Lignum Dalbergiae benthamii extract as claimed in claim 2, is characterized in that, during each reflux, extract, the consumption of ethyl acetate is 5-10 times of thing weight to be extracted.

6. Lignum Dalbergiae benthamii extract as claimed in claim 1 and the application as the Lignum Dalbergiae benthamii extract in claim 2-5 prepared by any one, is characterized in that, described Lignum Dalbergiae benthamii extract is preparing the application in antiinflammatory and antibacterial pharmaceutical preparation.

7. the application of Lignum Dalbergiae benthamii extract as claimed in claim 6, is characterized in that, described pharmaceutical preparation includes Lignum Dalbergiae benthamii extract and the pharmaceutically acceptable adjuvant composition of effective amount.

8. the application of Lignum Dalbergiae benthamii extract as claimed in claim 7, it is characterized in that, described pharmaceutical preparation is oil preparation, ointment, ointment, paste or plaster.